In situ hybridization study of cytokeratin 4, 13, 16 and 19 mRNAs in human developing junctional epithelium.
Cytokeratins (CKs) are now considered to be reliable markers for following the development and differentiation of epithelial tissue. We have investigated the pathway of differentiation in human developing junctional epithelium using monoclonal antibodies and two-dimensional gel electrophoresis of microdissected tissue to identify CK 19, CK 16, CK 14, CK 13, CK 6, CK 5, CK 4 in the junctional epithelium (JE) over partially erupted human teeth. The CK profile was similar to that of developing oral epithelia, suggesting that the junctional epithelium in teeth during eruption is of odontogenic origin. The present study used in situ hybridization to determine the distribution of the mRNAs of CKs 19, 16, 13 and 4 in human developing junctional epithelium and to examine the correlation between mRNAs and their encoded proteins. CK 19 mRNA was abundant in the basal cell layers of the primary junctional epithelium (PJE) but less concentrated in the suprabasal layers. CK16, 13 and 4 mRNAs were abundant in the basal cell layers of the PJE. The parabasal cell layers reacted intensely to the cRNA probe complementary to CK16 mRNA, as were the reactions in the suprabasal cell layers of the PJE for the CK 13 and 4 probes. Our results demonstrate that the PJE express the genes encoding for CKs 16 and 4 that have been revealed previously only by electrophoresis. They therefore confirm that the PJE is a well-differentiated stratified epithelium with a complex unique phenotype that produces CKs specific for basal cells (CK 19), CKs associated with hyperproliferation (CK 16), and finally those associated with stratification (CKs 4 and 13). Only synthesis of CK 19 protein and mRNA are strictly parallel. CKs 4 and 13 mRNAs are present in basal and suprasal cells, while their encoded proteins were not, except for CK 13 in suprabasal cell layers of PJE, where the amount of its mRNAs was coincident with the expression of the protein.
Feghali-Assaly
M
M
Laboratoire de Recherches Orthopédiques, URA CNRS 1432, Faculté de Médecine Lariboisière Saint-Louis, Paris, France.
Sawaf
M H
MH
Ouhayoun
J P
JP
D000293:Adolescent; D000911:Antibodies, Monoclonal; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation; D002455:Cell Division; D002648:Child; D004396:Coloring Agents; D015180:Electrophoresis, Gel, Two-Dimensional; D004846:Epithelial Attachment / Q000378:metabolism*; D004847:Epithelial Cells / Q000502:physiology; D004848:Epithelium / Q000254:growth & development; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007633:Keratins / Q000032:analysis* / Q000145:classification / Q000235:genetics; D009805:Odontogenesis; D010641:Phenotype; D015347:RNA Probes; D018075:RNA, Complementary; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D014078:Tooth Eruption
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D004396:Coloring Agents; D015347:RNA Probes; D018075:RNA, Complementary; D012333:RNA, Messenger; D007633:Keratins